Cushing D J, Baez M, Kursar J D, Schenck K, Cohen M L
Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285.
Life Sci. 1994;54(22):1671-80. doi: 10.1016/0024-3205(94)00607-5.
The identity of the serotonin (5-HT) receptor(s) that mediate(s) contraction in canine coronary artery and saphenous vein remains controversial. Ring segments of endothelium-denuded coronary artery and helical strips of saphenous vein were suspended in organ chambers for measurement of isometric force. 5-HT, alpha Me-5-HT and sumatriptan contracted both coronary artery and saphenous vein and the non-selective 5-HT receptor antagonist 1-naphthylpiperazine (100nM) blocked 5-HT- and sumatriptan-induced contraction in both tissues. The agonist rank order potency for contraction (5-HT > sumatriptan > alpha Me5-HT > 5-MeOT > 5-MeT) was similar in both tissues and was consistent with that for a 5-HT1D receptor. Oligonucleotide primers specific for the 5-HT1D receptor sequence were designed for use in a polymerase chain reaction (PCR). cDNA derived from total RNA or mRNA from canine tissues was used in the PCR. PCR resulted in the amplification of a 632 base pair sequence in both canine coronary artery and saphenous vein; consistent with that expected for the 5-HT1D receptor. Southern blot analysis, with an oligonucleotide probe internal to the sequence amplified by the PCR primers, confirmed that the sequence amplified by PCR was the 5-HT1D receptor. Thus, the 5-HT1D receptor is expressed in canine coronary artery and saphenous vein and taken together with the pharmacological data, supports the possibility that a 5-HT1D-like receptor mediates contraction in these two tissues.
介导犬冠状动脉和隐静脉收缩的5-羟色胺(5-HT)受体的身份仍存在争议。将去内皮冠状动脉的环形节段和隐静脉的螺旋条带悬挂在器官浴槽中以测量等长力。5-HT、α-甲基-5-HT和舒马曲坦使冠状动脉和隐静脉均收缩,非选择性5-HT受体拮抗剂1-萘基哌嗪(100nM)可阻断5-HT和舒马曲坦在这两种组织中诱导的收缩。两种组织中激动剂诱导收缩的效价顺序(5-HT>舒马曲坦>α-甲基-5-HT>5-甲氧基色胺>5-甲基色胺)相似,且与5-HT1D受体的顺序一致。设计了针对5-HT1D受体序列的寡核苷酸引物用于聚合酶链反应(PCR)。来自犬组织总RNA或mRNA的cDNA用于PCR。PCR在犬冠状动脉和隐静脉中均扩增出一条632个碱基对的序列;与5-HT1D受体预期的序列一致。用PCR引物扩增序列内部的寡核苷酸探针进行Southern印迹分析,证实PCR扩增的序列为5-HT1D受体。因此,5-HT1D受体在犬冠状动脉和隐静脉中表达,结合药理学数据,支持5-HT1D样受体介导这两种组织收缩的可能性。